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Insecticide Resistance Action Committee How to Develop an Insect Resistance Management Plan: Practical Approaches for Local Environments CLI-IRAC Presentation on Insect Resistance Management 2009 1 Today’s Topics Insecticide Resistance Action Committee Global Adoption of Agriculture Biotechnology – Characteristics of insect control traits Why is Insect Resistance Management (IRM) Important for Transgenic Insecticidal Crops? – Scientific theory of IRM strategy – Definitions & purpose – Foundation and tactics CLI-IRAC Presentation on Insect Resistance Management 2009 Today’s Topics Insecticide Resistance Action Committee USA Experience with Insect Resistance Management – Maize, Pyramiding Insecticidal Proteins, Cotton Global Experience with IRM How to Develop an IRM Plan CLI-IRAC Presentation on Insect Resistance Management 2009 3 Biotechnology in Agriculture Insecticide Resistance Action Committee Biotechnology has been enthusiastically embraced by growers around the world – More than 125M Ha were planted in 2007 – Grown by 13.3M growers in 25 different countries – 90% of these growers are in developing nations – The fastest growing regions are the developing nations CLI-IRAC Presentation on Insect Resistance Management 2009 4 Global Distribution of Biotech Crops Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 5 Insect Control Traits Insecticide Resistance Action Committee While most of these biotech hectares are planted with herbicide tolerant traits, about 33% of the hectares (>40M Ha) are planted with insect control traits, Bt and/or VIP Bacillus thuringiensis (Bt) a natural bacterium that produces insect control proteins, called Cry and VIP proteins – Cry = crystal proteins because they form crystals in the native bacterium and control coleopteran or lepidopteran crop pests – VIP = Vegetative insecticidal proteins are secreted during the vegetative bacterial stages prior sporulation CLI-IRAC Presentation on Insect Resistance Management 2009 6 Insecticide Resistance Action Committee Cry and VIP proteins have superior environmental and health benefits! CLI-IRAC Presentation on Insect Resistance Management 2009 7 Mode of Action of Bt Cry Proteins Insecticide Resistance Action Committee VIP’s also are proteolytically activated, bind to insect midgut cells, and form pores; however, VIP’s bind to independent receptors from Cry protein on the insect midgut. CLI-IRAC Presentation on Insect Resistance Management 2009 8 Resistance – What is it? Insecticide Resistance Action Committee Resistance is defined as the ability of an organism to ‘defend’ itself from disease. – In agriculture, resistance has a slightly different meaning, when a pest can defend itself from the controlling agent: When weeds are able to survive an herbicide When insects survive an insecticide When insect populations develop resistance, it is typically a gradual process where they are able to withstand higher and higher amounts of insecticide, until finally, the insecticide is no longer effective in controlling the insect pest. Not all insects develop resistance, pest biology and insecticide selection pressure influence the result CLI-IRAC Presentation on Insect Resistance Management 2009 9 Insect Resistance Management Insecticide Resistance Action Committee The greatest risks to Bt and VIP crops is the potential to develop insect resistance: Long-term product durability is the goal! Resistance has occurred with sprayable Bt sprayable formulations since the mid-1990s – Mostly Diamondback moth populations in tropical regions Highly effective insect control with Bt and VIP crops has led to concerns of insect resistance evolution CLI-IRAC Presentation on Insect Resistance Management 2009 10 Insect Resistance Management Insecticide Resistance Action Committee Utilize IRM knowledge and experience: >10 years of Bt crop success – Argentina, Australia, Canada, China, Philippines, United States BUT IRM plans must be tailored to local or regional environments! CLI-IRAC Presentation on Insect Resistance Management 2009 11 Resistance Management Basics Insecticide Resistance Action Committee Definitions: Susceptible insect = one readily killed by the toxin susceptible insect + Bt or VIP = Resistant insect = one that survives the toxin resistant insect + Bt or VIP = CLI-IRAC Presentation on Insect Resistance Management 2009 12 Definitions: Genetic Traits Insecticide Resistance Action Committee Allele = a pair of genes that cause a trait Genotype = the genetic make-up of an organism, the genotype cannot be seen Phenotype = the observable trait or properties of an organism Genotype Phenotype susceptible insect = s / s Alleles resistant insect = r/r CLI-IRAC Presentation on Insect Resistance Management 2009 Alleles 13 Definitions: Genetic Dominance Insecticide Resistance Action Committee Alleles can have different ‘strengths’, meaning that they can be dominant or recessive in their effect on the trait phenotype Dominant = a gene combination that effects the phenotype of an organism with a single allele Recessive = a genetic make-up that has no impact on the phenotype unless it present in both allele. CLI-IRAC Presentation on Insect Resistance Management 2009 14 Trait Dominance: Examples Insecticide Resistance Action Committee Genotype Phenotype Trait Dominance Recessive allele susceptible insect = s/s Alleles Unknown susceptible insect = r/s Alleles Recessive resistant insect = r/r Alleles Recessive susceptible insect = s / s Alleles Unknown resistant insect = s / R Alleles Dominant resistant insect = R / R Alleles Dominant Dominant allele s = susceptible allele, r = recessive resistance allele, R = dominant resistance allele CLI-IRAC Presentation on Insect Resistance Management 2009 15 Definition: Dose Insecticide Resistance Action Committee Dose = the amount of insecticidal protein in the plant: – High dose kills all target insects and all heterozygous insects (offspring of resistant and susceptible insects) – Medium dose kills close to 100% of the susceptible insects – Low dose kills significantly less than 100% of the susceptible insect CLI-IRAC Presentation on Insect Resistance Management 2009 16 Genetic Dominance & Dose Insecticide Resistance Action Committee If the trait is: Recessive OR Phenotype X= Dose Phenotype Dose Med/High 25% Susceptible Med/High Susceptible 50% Susceptible High Resistant 25% Resistant Parents Dominant Resistant Offspring Population (F1) percentage What part of the population was controlled? CLI-IRAC Presentation on Insect Resistance Management 2009 75% 25% 17 Genetic Dominance and Dose Insecticide Resistance Action Committee % Mortality High Dose Level s/s s/r s/R r/r Dose or Toxin Concentration CLI-IRAC Presentation on Insect Resistance Management 2009 18 Dose & Susceptibility Insecticide Resistance Action Committee As larvae grow, they become less sensitive to the insecticidal protein and will cause more feeding damage This effect is unrelated to resistance. CLI-IRAC Presentation on Insect Resistance Management 2009 19 Resistance Management Basics Insecticide Resistance Action Committee Purpose: An Insect Resistance Management (IRM) Plan utilizes various activities to minimize selection pressure on the target pests. IRM reduces pest populations by as many different tactics as possible to maintain the ability to control the target pests: Analogous to Integrated Pest Management. Other aspects communicate the plan to stakeholders, monitor the effectiveness of the IRM plan and ensure compliance with it’s elements. To extend the product durability for as long as possible CLI-IRAC Presentation on Insect Resistance Management 2009 Foundation of an IRM Plan Insecticide Resistance Action Committee The biology of the crop, pest and crop/pest interaction is the foundation of an IRM plan – Crop biology will differ according to the growing region – The pest spectrum will also vary – The insect movement patterns of both larvae and adults will be unique for each crop The target pest susceptibility to the Cry or VIP protein is determined to shape the IRM plan – Is the expression level in the insecticidal crop sufficient to control the pest(s)? – Is the level considered low, medium or high dose? CLI-IRAC Presentation on Insect Resistance Management 2009 21 Experience with IRM Tactics Insecticide Resistance Action Committee Initial IRM plans were based on: – Crops expressing a ‘high dose’ of the Cry or VIP proteins High enough to kill both susceptible insects and heterozygous offspring when resistance trait is recessive – A structured refuge, planted close to or within the insecticidal crop, to produce large numbers of susceptible insects – Monitoring of the insecticidal crop to look for higher than expected levels of damage that might indicate development of resistant insect populations – The assumption that resistance would be functionally recessive and rare in insect populations CLI-IRAC Presentation on Insect Resistance Management 2009 22 In theory… Insecticide Resistance Action Committee Insect resistance will be delayed, when resistant alleles are rare and when the refuge produces sufficient numbers of susceptible insects to vastly outnumber the rare resistant insects that might emerge from the insecticidal crop. CLI-IRAC Presentation on Insect Resistance Management 2009 23 Refuge Strategy: Recessive Trait Insecticide Resistance Action Committee = susceptible = resistant Non-Bt Refuge CLI-IRAC Presentation on Insect Resistance Management 2009 Bt or VIP Crop 24 IRM Logo for Bt Crops in USA Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 25 Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 An Example: Bt Maize in the USA 26 Insecticide Resistance Action Committee Refuge Deployment Options: Maize Bt or VIP field Non-Bt or non-VIP Block Strips CLI-IRAC Presentation on Insect Resistance Management 2009 Border 27 Insecticide Resistance Action Committee Pyramided Bt Crops & Improved IRM Plans Optimized IRM plans have been developed based on experience and research results Insecticidal crops that express 2 or more insecticidal proteins are now commercial: called pyramided crops – The Cry and/or VIP proteins control the same insect – Killing the insect by unique sites of action (binding sites) – Pyramided insecticidal crops have a lower risk of resistance Natural refuge can be an important source of insects, especially generalists – insects that feed on a wide variety of crop species – Natural refuge consists of all types of different non-crop plants that can serve as hosts for the target pest CLI-IRAC Presentation on Insect Resistance Management 2009 28 Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 An Example: Bt Cotton in USA 29 Refuge Deployment Options: Cotton Insecticide Resistance Action Committee 5% Non-Bt Non-crop Single gene Bt field Pyramided Bt or VIP field Single gene Bt field Non-Bt 20% External Block Embedded CLI-IRAC Presentation on Insect Resistance Management 2009 Natural Refuge 30 An Example: Pyramided Maize Products Insecticide Resistance Action Committee Pyramided maize products, MON89034 and Bt11xMIR162, have been granted reduced refuge in the US In cotton growing regions, both products have been granted a 20% non-Bt corn refuge (reduced from 50%) – Both products offer two modes of action vs H. zea Only MON89034 has been granted a reduced refuge in corn growing regions to 5% (reduced from 20%) – Bt11xMIR62 only offers one mode of action vs. O. nubilalis Deployment and Proximity requirements remain unchanged from single gene maize products CLI-IRAC Presentation on Insect Resistance Management 2009 31 Other IRM Plan Elements – USA Insecticide Resistance Action Committee Monitoring for susceptibility of the target pest(s) – Collect insect populations from greatest concentration of insecticidal crop areas or in association with crop damage – Routinely conduct insect bioassays using specialized screening techniques – Goal: Identify population shifts in sensitivity to the Bt and/or VIP active that indicate increasing risk of resistance occurring and need for proactive remedial action CLI-IRAC Presentation on Insect Resistance Management 2009 32 Bt Maize Growing Regions – USA Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 33 Resistance Monitoring Regions Central Corn Belt Insecticide Resistance Action Committee 6 1 3 2 5 23 30 4 42 24 13 1 18 1 41 16 23 47 3 20 8 17 5 29 8 21 48 34 7 17 15 11 16 37 12 2 22 2 19 4 19 20 33 4 27 18 35 43 28 24 20 26 31 26 6 45 3 44 39 8 29 46 6 1 10 17 22 25 36 28 15 22 9 11 25 10 14 5 27 2 40 32 38 13 30 12 7 14 31 4 55 41 35 5 4 2 7 3 6 16 30 19 8 57 32 38 46 10 61 26 14 22 51 5 27 25 47 34 1 23 29 21 64 2 16 1 20 10 18 7 15 5 9 8 6 4 15 4 17 16 21 5 12 1 8 3 9 7 6 7 13 16 84 87 61 41 85 3 9 18 31 21 55 66 26 28 6 13 51 12 37 16 30 18 5 3 33 9 32 77 65 19 68 1 29 20 10 19 23 8 1 1 42 14 88 31 11 69 17 10 70 10 3 36 9 6 9 24 27 31 67 76 2 42 17 83 11 15 86 43 74 12 36 38 34 4 14 2 7 45 4 54 2 48 63 60 53 49 10 14 32 31 62 22 13 33 7 78 59 79 5 21 4 7 14 39 58 13 6 74 38 14 15 39 34 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grower compliance of IRM plans – Is refuge deployed according to the plan? – Re-enforcing education, when growers are found out of compliance Remedial action plans when resistance is suspected and/or confirmed CLI-IRAC Presentation on Insect Resistance Management 2009 35 Experience vs. Local Needs Insecticide Resistance Action Committee While experience is important, the local environment is equally as important! THEREFORE, what has worked for large plantings of monoculture production systems in North America may not be effective for small, diverse agricultural farms found in other regions CLI-IRAC Presentation on Insect Resistance Management 2009 36 IRM Plans: Australia Insecticide Resistance Action Committee Initial IRM plans limited each grower to planting a maximum of 30% of his farm to Bt cotton – Specifically recognized the higher risk of Bt crops with a single Cry protein Planting times are restricted Limited insecticide use on refuges Require post-harvest cultivation to reduce overwintering populations – ‘Pupae busting’ Once pyramided Bt cotton was commercialized; the planting cap of 30% was lifted CLI-IRAC Presentation on Insect Resistance Management 2009 37 IRM Plans: China Insecticide Resistance Action Committee No structured refuge requirement – Growers farm plots of 5Ha or less – Mixtures of crop species are present – Thus abundant natural refuge is available Annual monitoring for insect susceptibility is conducted for all commonly used insecticides, including the Cry proteins in Bt cotton – Old-World Cotton Bollworm is primary target pest To date cotton (Bt and Bt/CpTI) is the only Bt crop registered for cultivation, limiting insect selection pressure CLI-IRAC Presentation on Insect Resistance Management 2009 38 IRM Plans: India Insecticide Resistance Action Committee Growers are requested to plant a non-Bt refuge around each plot of Bt cotton – Refuge must be at least 5 rows wide – The non-Bt refuge must be at least 20% of the total cotton plot The company supplies the non-Bt seed in a package attached to the Bt cotton seed – For each 450g Bt cotton seed, 120g non-Bt cotton seed is included Frequent monitoring of insect susceptibility is conducted CLI-IRAC Presentation on Insect Resistance Management 2009 39 How to Develop an IRM Plan Insecticide Resistance Action Committee Several elements are common to all IRM plans: Identify key stakeholders to participate in IRM plan development and maintenance – Scientific experts on crop and pest biology – Grower groups/advisors know the local production practices – Commodity groups and product developers can provide education and communication about the technology – Regulatory authorities can ensure compliance with IRM plans CLI-IRAC Presentation on Insect Resistance Management 2009 40 How to Develop an IRM Plan Insecticide Resistance Action Committee Information about crop biology, target pest biology and crop/pest interactions should be determined – Insect movement and feeding patterns are key – Does the Bt crop provide a high level of insect control? – Pyramided Cry and/or VIP proteins? Local growing conditions should be described – Are crops grown in large monocultures? – Or are mixtures of crops more prevalent? – How big is the typical farm size? – How will local or regional cultural practices impact IRM plans? CLI-IRAC Presentation on Insect Resistance Management 2009 41 How to Develop an IRM Plan Insecticide Resistance Action Committee Growers and all stakeholders should fully appreciate the insecticidal crop technology, the superior safety profile and how it is best deployed for long term durability – What pests will be controlled? – What are IRM plan tactics and requirements? Will there be… Refuges deployed? Limits on insecticidal crop sales? Monitoring market penetration to set action triggers? Cultural practices to limit insect selection pressure? – What are all the crop deployment options? CLI-IRAC Presentation on Insect Resistance Management 2009 42 How to Develop an IRM Plan Insecticide Resistance Action Committee The responsibilities should especially be clear to the grower through education and communication – Is there a maximum amount of insecticidal crop that can be planted at one farm? – Is a non-insecticidal refuge required? If so, how much? And where should the refuge be placed? – Can insecticides be used? On the insecticidal crop? On the non-insecticidal refuge? Both? If so, what is the economic threshold? – How does the grower report unexpected insect damage? – What records should be kept? CLI-IRAC Presentation on Insect Resistance Management 2009 43 How to Develop an IRM Plan Insecticide Resistance Action Committee Establish insect susceptibility baseline Establish routine monitoring plan for insect susceptibility and for grower compliance – Decide the frequency for all monitoring Monitoring for insect susceptibility – Develop a method for screening insect populations Discriminating dose assay is one method – Collect representative insect populations from areas where the highest amount of insecticidal crop is planted – Conduct routine insect and field monitoring plan CLI-IRAC Presentation on Insect Resistance Management 2009 44 How to Develop an IRM Plan Insecticide Resistance Action Committee Monitoring for grower compliance to IRM plan – Determine an unbiased method to assess compliance – When growers are found out of compliance, what are the consequences? Additional education or incentives? Remove access to insecticidal crop technology? Remedial action plans established before or soon after commercial launch CLI-IRAC Presentation on Insect Resistance Management 2009 45 Insecticide Resistance Action Committee Thank you for your attention! Are there any questions? CLI-IRAC Presentation on Insect Resistance Management 2009 46 Bt Maize Reduced Fungal Toxins Insecticide Resistance Action Committee Fungi can produce harmful toxins in our crops Mycotoxins cause acute illness and cancer in humans and animals – Insect damage on crops allows fungi to grow that produces mycotoxins – Because Bt maize reduces insect damage, fungal growth is also reduced Bt maize reduces mycotoxin levels to ½ of conventional maize Beneficial for animal and human health CLI-IRAC Presentation on Insect Resistance Management 2009 47 Mycotoxins in Maize Insecticide Resistance Action Committee Fumonisin Fusarium verticillioides, F. proliferatum Aflotoxin Aspergillus flavus, A. parasiticus Deoxynivalenol (DON, vomitoxin) F. graminearum, F. culmorum Zearalenone F. graminearum, F. culmorum Examples of Ear Rot Fusarium Gibberella CLI-IRAC Presentation on Insect Resistance Management 2009 Aspergillus Photos: Gary Munkvold 48 Results: Bt Maize & Fumonisin Insecticide Resistance Action Committee CLI-IRAC Presentation on Insect Resistance Management 2009 49